CN112174976A - Dibenzofuran-type lignans isolated from cress and their methods and use in anti-gouty arthritis - Google Patents
Dibenzofuran-type lignans isolated from cress and their methods and use in anti-gouty arthritis Download PDFInfo
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Abstract
从水芹菜中分离的二苯并呋喃型木脂素及其方法和在抗痛风性关节炎上的用途，本发明的提取工艺，以水芹菜为原料，综合采用乙醇回流提取，硅胶柱色谱、凝胶色谱和液相色谱制备等步骤协同配合的方式，从活性部位中成功分离得到了一种新二苯并呋喃型木脂素。提取得到的成品化合物结构新颖，提取方法简便快捷、易于操作、且产率较高。本发明还基于小鼠踝关节注射尿酸钠结晶建立痛风性关节炎模型，测定水芹菜中提取到的新二苯并呋喃型木脂素对小鼠血清中IL‑1β和TNF‑α水平的影响，评价其抗痛风性关节炎活性。首次公开了新二苯并呋喃型木脂素在制备抗痛风性关节炎药物方面的用途，为今后临床抗痛风性关节炎治疗药物的研究和开发奠定了基础。
Dibenzofuran-type lignans separated from water celery, method thereof and use in anti-gouty arthritis, the extraction process of the present invention uses water celery as raw material, comprehensively adopts ethanol reflux extraction, silica gel column chromatography, condensation A new dibenzofuran-type lignan was successfully isolated from the active site by the synergistic cooperation of gel chromatography and liquid chromatography preparation. The finished product compound obtained by extraction has novel structure, and the extraction method is simple, quick, easy to operate, and has high yield. The invention also establishes a gouty arthritis model based on the injection of sodium urate crystals into the ankle joint of mice, and determines the effect of the new dibenzofuran-type lignans extracted from cress on the levels of IL-1β and TNF-α in the serum of mice , to evaluate its anti-gouty arthritis activity. The use of new dibenzofuran-type lignans in the preparation of anti-gouty arthritis drugs is disclosed for the first time, which lays a foundation for the research and development of clinical anti-gouty arthritis drugs in the future.
Description
Technical Field
The invention relates to the technical field of medicines, in particular to a novel dibenzofuran lignan separated from water cress, a preparation method thereof and application of the compound in preparing anti-gouty arthritis medicines.
Background
Oenanthe stolonifera (B)Oenanthe Javanica) Also called cress, etc., generally grow on low-lying wetlands, shallow water marshes or river banks. The perennial root herbaceous aquatic plant belongs to an umbelliferae, has a long and thin stem, is hollow, has creeping property, can root among all internodes to form a new plant, and is a traditional characteristic aquatic vegetable in China. The medicinal use of the water cress has a long history, and the historical documents record that the water cress has the effects of clearing heat, promoting diuresis, reducing phlegm, descending qi, removing blood stasis, stopping leukorrhagia, detoxifying, reducing swelling, clearing liver, benefiting gallbladder and the like, and is mainly used for treating symptoms such as jaundice, dispelling wind and the like.
In recent years, the chemical components of the cress are actively researched at home and abroad, and the main chemical components of the cress comprise flavone, alkaloid, coumarin, volatile oil, sugar, fatty acid, protein, amino acid and the like. Modern pharmacological research finds that the cress also has the effects of reducing blood pressure, blood fat and blood sugar, resisting cancer, protecting nerves, resisting hepatitis and the like. In order to further develop and utilize the rich natural resource of the water cresses, the invention adopts a method of biological activity to guide separation, separates a new dibenzofuran lignan for treating gouty arthritis from the water cresses, and documents and patent reports related to the extraction of the new dibenzofuran lignan from the water cresses and the activity are not seen at home and abroad so far.
Disclosure of Invention
The technical purpose of the invention is as follows: the compound is prepared from medicinal and edible water cress as a raw material, is separated and purified from the water cress by a process method with simple process and convenient extraction, and provides the application of the compound in preparing medicines for resisting gouty arthritis.
In order to realize the purpose, the invention adopts the following technical scheme: dibenzofuran-type lignans isolated from Oenanthe javanica, having a chemical name of: 2-propenyl-3, 4- (9-isopropanoxy-furan) -5-methoxy-5 ' - (4 "-hydroxy-3", 5 "-dimethoxyphenyl) -4' -methanol-2 ', 3' - (11',12' -dimethyl-8 ',9' -dicarbonyl) -1,1' -dibenzofuran type lignan with molecular formula C35H34O10The structural formula is as follows:
a method for separating dibenzofuran lignans from water cress comprises the following steps:
(1) pulverizing dried herba Apii Graveolentis into coarse granules, extracting with 70-90% ethanol under heating for several times under reflux, and mixing the extractive solutions;
(2) concentrating and drying the extracting solution prepared in the step (1) under the vacuum condition at the temperature of 40-60 ℃ to prepare dry total thick paste for later use;
(3) adding the total thick paste prepared in the step (2) into distilled water according to the volume ratio of 1 (0.9-1.3), ultrasonically dispersing to prepare a dispersion liquid in which the total thick paste is uniformly suspended in the water, then sequentially extracting the dispersion liquid by using cyclohexane, ethyl acetate and n-butyl alcohol, and performing rotary evaporation drying on the obtained extraction liquid of each part to obtain a cyclohexane part, an ethyl acetate part and an n-butyl alcohol part respectively for later use;
(4) and (3) applying the ethyl acetate part obtained in the step (3) to a silica gel column, and then sequentially adopting a solvent with the volume ratio of 12: 1-3: the petroleum ether-ethyl acetate elution system of 1 carries out gradient elution to respectively obtain A, B, C, D fractions which are totally 4 fractions;
(5) and (3) applying the fraction D obtained in the step (4) to a silica gel column, and then sequentially adopting a volume ratio of 4: 1-2: the petroleum ether-ethyl acetate elution system of 1 carries out gradient elution to respectively obtain 3 sub-fractions of D-1, D-2 and D-3;
(6) putting the sub-fraction D-1 obtained in the step (5) on a Sephadex LH-20 gel column, then eluting by adopting a MeOH solution with the mass concentration of 95%, and obtaining 3 sub-fractions of D-1-1, D-1-2 and D-1-3 according to the color of a color band after elution;
(7) and (3) repeatedly putting the sub-fraction D-1-2 obtained in the step (6) on A silicA gel column and A Sephadex LH-20 gel column for separation and purification, then preparing A liquid phase YMC-Pack ODS-A column, and separating and purifying to obtain the light yellow solid dibenzofuran lignans.
Further, in the step (1), the particle size of the crude particles obtained after the water cress is crushed is 70-100 meshes.
Further, in the step (1), the times of reflux extraction are 3-5 times, and the time of each reflux extraction is 4 h.
Further, in the step (4), the particle size of the filler in the silica gel column is 100-200 meshes.
Further, in the step (5), the particle size of the filler in the silica gel column is 200-300 meshes.
Further, in the step (7), the chromatographic conditions of the liquid phase column are as follows: absorption wavelength: 202-350 nm, mobile phase: 15-40% methanol.
Further, in step (7), the liquid phase YMC-Pack ODS-A column has A size of 250X 10 mm, S-5 μm, 12 nm.
The dibenzofuran lignan separated from the water cress is applied to preparing medicaments for resisting gouty arthritis.
The invention has the beneficial effects that:
the extraction process of the invention takes the water cress which is a vegetable used as both medicine and food as a raw material, and successfully separates a new dibenzofuran lignan from active sites by comprehensively adopting a mode of synergistic combination of the steps of ethanol reflux extraction, silica gel column chromatography, gel chromatography, liquid chromatography preparation and the like. The extracted finished product compound has novel structure, simple, convenient and quick extraction method, easy operation and higher yield. The invention also establishes a gouty arthritis model based on the injection of sodium urate crystal to the ankle joint of the mouse, measures the influence of the new dibenzofuran lignans extracted from the cress on the IL-1 beta and TNF-alpha levels in the serum of the mouse, and evaluates the anti-gouty arthritis activity of the new dibenzofuran lignans. The application of the novel dibenzofuran lignan in preparing the anti-gouty arthritis medicament is disclosed for the first time, and a foundation is laid for the research and development of clinical anti-gouty arthritis treatment medicaments in the future.
Drawings
FIG. 1 is a flow chart of a novel monomer compound I prepared in example 1 of the present invention;
FIG. 2 is a diagram showing the coupling of the novel monomeric compound I prepared in example 1 of the present invention.
Detailed Description
The present invention will be described in further detail with reference to examples, but the scope of the present invention is not limited thereto.
The invention mainly provides a novel dibenzofuran lignan separated from water cress, a preparation method of the compound and anti-gouty arthritis activity. The new compound is separated from the water cress.
The name of the new monomer compound I is: 2-propenyl-3, 4- (9-isopropanoxy-furan) -5-methoxy-5 ' - (4 "-hydroxy-3", 5 "-dimethoxyphenyl) -4' -methanol-2 ', 3' - (11',12' -dimethyl-8 ',9' -dicarbonyl) -1,1' -dibenzofuran type lignan
Novel monomeric compounds of formula i: c35H34O10
Novel monomeric compounds of formula i:
in order to obtain the novel monomer compound I, the preparation method comprises the following steps:
(1) pulverizing dried herba Apii to obtain crude product (70-100 mesh), and extracting with 70-90% ethanol under reflux for 3-5 times (4 hr/time) to obtain total extractive solution;
(2) concentrating and drying the extracting solution obtained in the step (1) under the vacuum condition of low temperature (40-60 ℃) to obtain dry total thick paste;
(3) uniformly suspending the total thick paste obtained in the step (2) in water (volume ratio =1:0.9-1.3, V/V), sequentially extracting with cyclohexane, ethyl acetate and n-butanol, and performing rotary evaporation drying on the obtained extraction liquid of each part to obtain a cyclohexane part, an ethyl acetate part and an n-butanol part respectively for later use;
(4) establishing a gouty arthritis model based on the injection of sodium urate crystals to ankle joints of mice by adopting a biological activity guidance separation method, measuring the influence of different extraction parts of the cress on the IL-1 beta and TNF-alpha levels in serum of the mice, and determining the ethyl acetate part in the step (3) as an anti-gouty arthritis active part;
(5) and (3) performing gradient elution on the ethyl acetate part obtained in the step (4) by using a silica gel (100-200 meshes) column and a petroleum ether-ethyl acetate elution system (12: 1 → 3: 1, V/V) to obtain 4 fractions A, B, C, D. Screening anti-gouty arthritis active sites of the 4 fractions, and determining the D fraction as an anti-gouty arthritis active fraction;
(6) applying the active fraction D obtained in the step (5) to a silica gel (200-300 mesh) column, and performing gradient elution by using a petroleum ether-ethyl acetate elution system (4: 1 → 2: 1) to obtain sub-fractions D-1, D-2 and D-3; screening anti-gouty arthritis active sites of the 3 sub-fractions, and determining that the sub-D-1 fraction is an anti-gouty arthritis active fraction;
(7) putting the sub-fraction D-1 obtained in the step (6) on a Sephadex LH-20 gel column, eluting with 95% MeOH, and obtaining sub-fractions D-1-1, D-1-2 and D-1-3 according to color band colors after elution; screening anti-gouty arthritis active sites of the 3 sub-fractions, and determining that D-1-2 is anti-gouty arthritis active sub-fraction;
(8) and (3) repeatedly applying the sub-fraction D-1-2 obtained in the step (7) to A silicA gel column and A gel column for separation and purification, and finally obtaining the new monomer compound I-dibenzofuran lignan in A light yellow solid through preparative liquid phase separation (wavelength: 202-350 nm, mobile phase: 15% -40% methanol, YMC-Pack ODS-A column: 250 x 10 mm, S-5 mu m and 12 nm).
Example 1:
preparation of novel monomeric compound i:
(1) pulverizing dried herba Apii (20.0 kg) into crude product (90 mesh), and extracting with 85% ethanol under reflux for 3 times (4 hr/time) to obtain total extractive solution;
(2) concentrating and drying the extracting solution obtained in the step (1) under the vacuum condition at low temperature (50 ℃) to obtain dry total thick paste (1.9 kg);
(3) uniformly suspending the total thick paste obtained in the step (2) in water (volume ratio =1:1, V/V), sequentially extracting with cyclohexane, ethyl acetate and n-butanol, and performing rotary evaporation drying on the obtained extraction liquid of each part to obtain a cyclohexane extraction part (79.4 g), an ethyl acetate extraction part (298.6 g) and an n-butanol extraction part (462.5 g) respectively;
(4) establishing a gouty arthritis model based on the injection of sodium urate crystals to ankle joints of mice by adopting a biological activity guidance separation method, measuring the influence of different extraction parts of the cress on the IL-1 beta and TNF-alpha levels in serum of the mice, and determining an ethyl acetate extraction part as an anti-gouty arthritis active part;
(5) the ethyl acetate fraction obtained in step (4) was subjected to silica gel (100-mesh 200-mesh) column and gradient elution with a petroleum ether-ethyl acetate elution system (12: 1 → 3: 1, V/V) to obtain 4 fractions A (40.2 g), B (61.6 g), C (75.9 g) and D (56.5 g). Screening anti-gouty arthritis active sites of the 4 fractions, and determining the D fraction as an anti-gouty arthritis active fraction;
(6) subjecting the active fraction D obtained in the step (5) to silica gel (200-300 mesh) column, and performing gradient elution with petroleum ether-ethyl acetate elution system (4: 1 → 2: 1) to obtain sub-fractions D-1 (13.5 g), D-2 (20.2 g) and D-3 (11.7 g); screening anti-gouty arthritis active sites of the 3 sub-fractions, and determining that the sub-D-1 fraction is an anti-gouty arthritis active fraction;
(7) putting the sub-fraction D-1 obtained in the step (6) on a Sephadex LH-20 gel column, eluting with 95% MeOH, and obtaining sub-fractions D-1-1 (4.1 g), D-1-2 (5.5 g) and D-1-3 (3.2 g) according to color band colors after elution; screening anti-gouty arthritis active sites of the 3 sub-fractions, and determining that D-1-2 is anti-gouty arthritis active sub-fraction;
(8) the sub-fraction D-1-2 obtained in step (7) was subjected to separation and purification by repeating the application to A silicA gel column and A gel column, and subjected to preparative liquid phase separation (wavelength: 210 nm, mobile phase: 28% methanol, YMC-Pack ODS-A column: 250X 10 mm, S-5 μm, 12 nm) to finally obtain A novel monomeric compound I (8.24 mg).
The new monomer compound I prepared in this example is light yellow solid, HR-ESI-MS m/z 637.2053[M+Na]+Indicating that its molecular formula is C35H34O10（calcd. for C35H34O10Na, 637.2050). UV (MeOH) of Compound Iλ max: 202、210、280 nm；IRν max: 3318、1660、1650、1525、1383 cm-1；1H NMR（CD3COCD3400 MHz) and13C NMR（CD3COCD3100 MHz) data are shown in table 1 below. From the spectral data of the novel monomeric compounds I and HMBC and1H-1the related information of H COSY coupling (figure 2) is searched by SciFinder to identify the compound I as a novel dibenzofuran lignan.
TABLE 1 preparation of compound I1H NMR(400 MHz, CD3COCD3)、13C NMR(100 MHz, CD3COCD3) And HMBC related data
Example 2:
preparation of novel monomeric compound i:
(1) pulverizing dried herba Apii (20.0 kg) into crude product (100 mesh), and extracting with 90% ethanol under reflux for 4 times (4 hr/time) to obtain total extractive solution;
(2) concentrating and drying the extracting solution obtained in the step (1) under the vacuum condition at low temperature (60 ℃) to obtain dry total thick paste (2.1 kg);
(3) uniformly suspending the total thick paste obtained in the step (2) in water (volume ratio =1:0.9, V/V), sequentially extracting with cyclohexane, ethyl acetate and n-butanol, and performing rotary evaporation drying on the obtained extract liquor of each part to obtain a cyclohexane extraction part (83.42 g), an ethyl acetate extraction part (302.4 g) and an n-butanol extraction part (453.9 g) respectively;
(4) establishing a gouty arthritis model based on the injection of sodium urate crystals to ankle joints of mice by adopting a biological activity guidance separation method, measuring the influence of different extraction parts of the cress on the IL-1 beta and TNF-alpha levels in serum of the mice, and determining an ethyl acetate extraction part as an anti-gouty arthritis active part;
(5) the ethyl acetate fraction obtained in step (4) was subjected to silica gel (100-mesh 200-mesh) column and gradient elution with a petroleum ether-ethyl acetate elution system (12: 1 → 3: 1, V/V) to obtain 4 fractions A (36.8 g), B (66.2.6 g), C (72.5 g) and D (60.1 g). Screening anti-gouty arthritis active sites of the 4 fractions, and determining the D fraction as an anti-gouty arthritis active fraction;
(6) subjecting the active fraction D obtained in the step (5) to silica gel (200-300 mesh) column, and performing gradient elution with petroleum ether-ethyl acetate elution system (4: 1 → 2: 1) to obtain sub-fractions D-1 (15.1 g), D-2 (19.6 g) and D-3 (12.5 g); screening anti-gouty arthritis active sites of the 3 sub-fractions, and determining that the sub-D-1 fraction is an anti-gouty arthritis active fraction;
(7) putting the sub-fraction D-1 obtained in the step (6) on a Sephadex LH-20 gel column, eluting with 95% MeOH, and obtaining sub-fractions D-1-1 (3.9 g), D-1-2 (5.8 g) and D-1-3 (3.9 g) according to color band colors after elution; screening anti-gouty arthritis active sites of the 3 sub-fractions, and determining that D-1-2 is anti-gouty arthritis active sub-fraction;
(8) the sub-fraction D-1-2 obtained in step (7) was subjected to separation and purification by repeating the application to A silicA gel column and A gel column, and subjected to preparative liquid phase separation (wavelength: 210 nm, mobile phase: 28% methanol, YMC-Pack ODS-A column: 250X 10 mm, S-5 μm, 12 nm) to finally obtain A novel monomeric compound I (8.31 mg).
Example 3:
preparation of novel monomeric compound i:
(1) pulverizing dried herba Apii (20.0 kg) into crude product (70 mesh), and extracting with 70% ethanol under reflux for 5 times (4 hr/time) to obtain total extractive solution;
(2) concentrating and drying the extracting solution obtained in the step (1) under the vacuum condition at low temperature (40 ℃) to obtain dry total thick paste (1.7 kg);
(3) uniformly suspending the total thick paste obtained in the step (2) in water (volume ratio =1:1.3, V/V), sequentially extracting with cyclohexane, ethyl acetate and n-butanol, and performing rotary evaporation drying on the obtained extract liquor of each part to obtain a cyclohexane extraction part (77.6 g), an ethyl acetate extraction part (300.1 g) and an n-butanol extraction part (466.4 g) respectively;
(4) establishing a gouty arthritis model based on the injection of sodium urate crystals to ankle joints of mice by adopting a biological activity guidance separation method, measuring the influence of different extraction parts of the cress on the IL-1 beta and TNF-alpha levels in serum of the mice, and determining an ethyl acetate extraction part as an anti-gouty arthritis active part;
(5) the ethyl acetate fraction obtained in step (4) was subjected to silica gel (100-mesh 200-mesh) column and gradient elution with a petroleum ether-ethyl acetate elution system (12: 1 → 3: 1, V/V) to obtain 4 fractions A (41.3 g), B (60.6 g), C (76.8 g) and D (55.4 g). Screening anti-gouty arthritis active sites of the 4 fractions, and determining the D fraction as an anti-gouty arthritis active fraction;
(6) subjecting the active fraction D obtained in the step (5) to silica gel (200-300 mesh) column, and performing gradient elution with petroleum ether-ethyl acetate elution system (4: 1 → 2: 1) to obtain sub-fractions D-1 (14.8 g), D-2 (22.1 g) and D-3 (10.9 g); screening anti-gouty arthritis active sites of the 3 sub-fractions, and determining that the sub-D-1 fraction is an anti-gouty arthritis active fraction;
(7) putting the sub-fraction D-1 obtained in the step (6) on a Sephadex LH-20 gel column, eluting with 95% MeOH, and obtaining sub-fractions D-1-1 (4.3 g), D-1-2 (6.1 g) and D-1-3 (3.4 g) according to color band colors after elution; screening anti-gouty arthritis active sites of the 3 sub-fractions, and determining that D-1-2 is anti-gouty arthritis active sub-fraction;
(8) the sub-fraction D-1-2 obtained in step (7) was subjected to separation and purification by repeating the application to A silicA gel column and A gel column, and subjected to preparative liquid phase separation (wavelength: 210 nm, mobile phase: 28% methanol, YMC-Pack ODS-A column: 250X 10 mm, S-5 μm, 12 nm) to finally obtain A novel monomeric compound I (8.12 mg).
The compound I prepared in the example 1 of the invention is subjected to an anti-gouty arthritis activity screening experiment study:
(1) experimental Material
SPF grade mice; an IL-1 beta ELISA kit; TNF-alpha ELISA kit; an enzyme-labeling instrument; a biological microscope; a high speed refrigerated centrifuge; CMC-Na solution (0.5%); MSU solution (3 mg/m L).
(2) Molding and administration
Mice were randomly divided into blank, model, positive, new monomeric compound i and labeled. The blank group and the model group were intragastrically administered with 0.5% CMC-Na solution, and the other groups were administered with the corresponding dose for 5 days in a continuous administration volume of 0.1 m L/10 g. After 3 days of administration, the model was made by bending the mouse posterior ankle to a right angle to sufficiently expose the gap between the ankle and the tibiofibula, and after local sterilization, the needle was inserted at 45 ° from the space between the two apophysis to make a sense of breakthrough, thereby proving that the needle entered the ankle. The blank group was injected with an equal amount of physiological saline, and the other administration groups except the blank group were injected with 50. mu.L of MSU solution. After the MSU solution is injected, the posterior ankle joint on the side of the model is observed, and if obvious swelling exists, the success of the model is indicated.
(3) Sample collection and processing
And (5) taking blood from the orbit after molding for 9 h, taking the blood with the amount of more than 0.1 m L, and placing in an ice water bath. Centrifuging at 4000 r/min for 5 min, collecting supernatant, and detecting IL-1 beta and TNF-alpha protein levels in serum according to IL-1 beta kit method. The experimental data are expressed as Mean ± standard error (Mean ± SE) and analyzed by SPSS 17.0, with an analysis of variance of F > 0.05 by the analysis of variance. And (3) performing two-tailed t test comparison between groups, and performing one-factor analysis of variance (LSD) test to judge whether the multiple groups have statistical significance, wherein P is less than 0.05, and the difference has statistical significance.
(4) Results of the experiment
The experimental result shows that the IL-1 beta concentration and the TNF-alpha concentration in the serum of the mouse in the model group are both obviously increased (P is less than 0.001, and P is less than 0.001). Compared with the model group, the IL-1 beta level of the positive group and the new monomer compound I is obviously reduced (P <0.001, P < 0.01); in addition, the positive group and the new monomer compound I can obviously inhibit the release of inflammatory factors TNF-alpha (P <0.001, P <0.05) (Table 2). Therefore, the novel monomer compound I has certain anti-gouty arthritis activity. The method lays an important theoretical foundation for the new monomer compound I in the aspect of preparing the anti-gouty arthritis medicine.
TABLE 2 Effect of novel monomeric Compound I on IL-1 beta and TNF-alpha levels in mouse serum
Note: in comparison with the blank set, the results,###p is less than 0.001; compared with the model groupp＜0.05，**p＜0.01，***p＜0.001。
Claims (9)
1. Dibenzofuran-type lignans isolated from Oenanthe javanica, characterized in that the chemical name of the dibenzofuran-type lignans is: 2-propenyl-3, 4- (9-isopropanoxy-furan) -5-methoxy-5 ' - (4 "-hydroxy-3", 5 "-dimethoxyphenyl) -4' -methanol-2 ', 3' - (11',12' -dimethyl-8 ',9' -dicarbonyl) -1,1' -dibenzofuran type lignan with molecular formula C35H34O10The structural formula is as follows:
2. a method for separating dibenzofuran lignans from water cress is characterized by comprising the following steps:
(1) pulverizing dried herba Apii Graveolentis into coarse granules, extracting with 70-90% ethanol under heating for several times under reflux, and mixing the extractive solutions;
(2) concentrating and drying the extracting solution prepared in the step (1) under the vacuum condition at the temperature of 40-60 ℃ to prepare dry total thick paste for later use;
(3) adding the total thick paste prepared in the step (2) into distilled water according to the volume ratio of 1 (0.9-1.3), ultrasonically dispersing to prepare a dispersion liquid in which the total thick paste is uniformly suspended in the water, then sequentially extracting the dispersion liquid by using cyclohexane, ethyl acetate and n-butyl alcohol, and performing rotary evaporation drying on the obtained extraction liquid of each part to obtain a cyclohexane part, an ethyl acetate part and an n-butyl alcohol part respectively for later use;
(4) and (3) applying the ethyl acetate part obtained in the step (3) to a silica gel column, and then sequentially adopting a solvent with the volume ratio of 12: 1-3: the petroleum ether-ethyl acetate elution system of 1 carries out gradient elution to respectively obtain A, B, C, D fractions which are totally 4 fractions;
(5) and (3) applying the fraction D obtained in the step (4) to a silica gel column, and then sequentially adopting a volume ratio of 4: 1-2: the petroleum ether-ethyl acetate elution system of 1 carries out gradient elution to respectively obtain 3 sub-fractions of D-1, D-2 and D-3;
(6) putting the sub-fraction D-1 obtained in the step (5) on a Sephadex LH-20 gel column, then eluting by adopting a MeOH solution with the mass concentration of 95%, and obtaining 3 sub-fractions of D-1-1, D-1-2 and D-1-3 according to the color of a color band after elution;
(7) and (3) repeatedly putting the sub-fraction D-1-2 obtained in the step (6) on A silicA gel column and A Sephadex LH-20 gel column for separation and purification, then preparing A liquid phase YMC-Pack ODS-A column, and separating and purifying to obtain the light yellow solid dibenzofuran lignans.
3. The method for separating dibenzofuran-type lignan from Oenanthe javanica according to claim 1, wherein: in the step (1), the particle size of the crude particles obtained after the water cress is crushed is 70-100 meshes.
4. The method for separating dibenzofuran-type lignan from Oenanthe javanica according to claim 1, wherein: in the step (1), the times of reflux extraction are 3-5 times, and the time of each reflux extraction is 4 h.
5. The method for separating dibenzofuran-type lignans from Oenanthe javanica as claimed in claim 1, wherein: in the step (4), the particle size of the filler in the silica gel column is 100-200 meshes.
6. The method for separating dibenzofuran-type lignans from Oenanthe javanica as claimed in claim 1, wherein: in the step (5), the particle size of the filler in the silica gel column is 200-300 meshes.
7. The method for separating dibenzofuran-type lignans from Oenanthe javanica as claimed in claim 1, wherein: in step (7), the chromatographic conditions of the liquid phase column are: absorption wavelength: 202-350 nm, mobile phase: 15-40% methanol.
8. The method for separating dibenzofuran-type lignans from Oenanthe javanica as claimed in claim 1, wherein: in step (7), the liquid phase YMC-Pack ODS-A column was 250X 10 mm in size, S-5 μm, 12 nm.
9. Use of the dibenzofuran-type lignan isolated from cress as set forth in claim 1 for the preparation of a medicament for treating gouty arthritis.
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